Autophagy-Related Gene PlATG6a Is Involved in Mycelial Growth, Asexual Reproduction and Tolerance to Salt and Oxidative Stresses in Peronophythora litchii.

Autophagy is ubiquitously present in eukaryotes. During this process, intracellular proteins and some waste organelles are transported into lysosomes or vacuoles for degradation, which can be reused by the cell to guarantee normal cellular metabolism. However, the function of autophagy-related (ATG) proteins in oomycetes is rarely known. In this study, we identified an autophagy-related gene, PlATG6a, encoding a 514-amino-acid protein in Peronophythora litchii, which is the most destructive pathogen of litchi. The transcriptional level of PlATG6a was relatively higher in mycelium, sporangia, zoospores and cysts. We generated PlATG6a knockout mutants using CRISPR/Cas9 technology. The P. litchii Δplatg6a mutants were significantly impaired in autophagy and vegetative growth. We further found that the Δplatg6a mutants displayed decreased branches of sporangiophore, leading to impaired sporangium production. PlATG6a is also involved in resistance to oxidative and salt stresses, but not in sexual reproduction. The transcription of peroxidase-encoding genes was down-regulated in Δplatg6a mutants, which is likely responsible for hypersensitivity to oxidative stress. Compared with the wild-type strain, the Δplatg6a mutants showed reduced virulence when inoculated on the litchi leaves using mycelia plugs. Overall, these results suggest a critical role for PlATG6a in autophagy, vegetative growth, sporangium production, sporangiophore development, zoospore release, pathogenesis and tolerance to salt and oxidative stresses in P. litchii.

The preference choices of Conopomorpha sinensis Bradley (Lepidoptera: Gracilariidae) for litchi based on its host surface characteristics and volatiles.

Conopomorpha sinensis Bradley is a host-specific pest of Litchi chinensis and Euphoria longan. Here, we demonstrated that C. sinensis has evolved special physical and chemical mechanisms for host plant location that enable it to survive and reproduce. Females favored laying their eggs on the convex surface of litchi fruit that had particular volatile characteristics. Experiments using a H-type olfactometer showed that female C. sinensis were attracted to litchi flowers, tender shoots, immature fruits, and mature fruits, with the highest attraction rate to mature fruits (74.67 ± 2.31%). There were no significant differences in the attraction of male C. sinensis to different litchi tissues. Further oviposition preference tests using the pericarp, pulp, and seeds of mature litchi fruits revealed that female C. sinensis prefer to lay their eggs on the pericarp. Litchi volatiles were found to be important in attracting C. sinensis to fruits for oviposition. Analysis of volatiles from different litchi tissues by HS-SPME-GC-MS revealed 31 similar volatiles, some of which may be important in the oviposition preference choices of C. sinensis on litchi fruit.

Sequencing of the Litchi Downy Blight Pathogen Reveals It Is a Phytophthora Species With Downy Mildew-Like Characteristics.

On the basis of its downy mildew-like morphology, the litchi downy blight pathogen was previously named Peronophythora litchii. Recently, however, it was proposed to transfer this pathogen to Phytophthora clade 4. To better characterize this unusual oomycete species and important fruit pathogen, we obtained the genome sequence of Phytophthora litchii and compared it to those from other oomycete species. P. litchii has a small genome with tightly spaced genes. On the basis of a multilocus phylogenetic analysis, the placement of P. litchii in the genus Phytophthora is strongly supported. Effector proteins predicted included 245 RxLR, 30 necrosis-and-ethylene-inducing protein-like, and 14 crinkler proteins. The typical motifs, phylogenies, and activities of these effectors were typical for a Phytophthora species. However, like the genome features of the analyzed downy mildews, P. litchii exhibited a streamlined genome with a relatively small number of genes in both core and species-specific protein families. The low GC content and slight codon preferences of P. litchii sequences were similar to those of the analyzed downy mildews and a subset of Phytophthora species. Taken together, these observations suggest that P. litchii is a Phytophthora pathogen that is in the process of acquiring downy mildew-like genomic and morphological features. Thus P. litchii may provide a novel model for investigating morphological development and genomic adaptation in oomycete pathogens.

Attraction of the redbay ambrosia beetle, Xyleborus glabratus, to avocado, lychee, and essential oil Lures.

The redbay ambrosia beetle, Xyleborus glabratus Eichhoff, is an exotic wood-boring insect that vectors the mycopathogen responsible for laurel wilt, a lethal vascular disease of trees in the Lauraceae. High mortality has occurred in native Persea species in the southeastern U.S., and the vector-pathogen complex poses an imminent threat to the production of commercial avocado, P. americana, in south Florida. There is a critical need for effective attractants to detect, monitor, and control this invasive pest. This study combined field tests and laboratory bioassays to evaluate the response of female X. glabratus to host-based volatiles from wood of avocado (cultivars of West Indian, Guatemalan, and Mexican races); from wood of lychee (Litchi chinensis, a presumed non-host that is high in the sesquiterpene α-copaene, a putative attractant); and to commercial lures containing manuka and phoebe oils, two reported attractive baits. Volatile collections and GC-MS analyses were performed to quantify the sesquiterpene content of test substrates. In the field, traps baited with lychee wood captured more beetles than those with wood from avocado cultivars; traps baited with phoebe oil lures captured more beetles than those with manuka oil lures (the current monitoring tool). In field and laboratory tests, X. glabratus did not show a preference among avocado races in either attraction or host acceptance (initiation of boring). In choice tests, lychee was more attractive than avocado initially, but a higher percentage of beetles bored into avocado, suggesting that lychee emits more powerful olfactory/visual cues, but that avocado contains more of the secondary cues necessary for host recognition. Emissions of α-copaene, ß-caryophyllene, and α-humulene were correlated with field captures, and lychee wood may be a source of additional semiochemicals for X. glabratus.

Entomopathogens (Beauveria bassiana and Steinernema carpocapsae) for biological control of bark-feeding moth Indarbela dea on field-infested litchi trees.

BACKGROUND: The bark-feeding moth Indarbela dea (Swinhoe) is one of the major pests of litchi (Litchi chinensis Sonn.) in Asia. Effects on larval mortality and web production were tested by repeated, single and combined tree trunk applications of the entomopathogenic fungus Beauveria bassiana (Bals.) Vuill. and the entomopathogenic nematode Steinernema carpocapsae (Weiser). Two separate application trials were conducted in different seasons on young and older larvae respectively. RESULTS: In both trials, complete or nearly complete mortality (100% and 95%) of the larvae was achieved 3 weeks after application of S. carpocapsae alone and 2 weeks after the combined application of B. bassiana and S. carpocapsae (100% in both trials). However, in the combined application, all larvae were killed by one treatment, while the application of S. carpocapsae alone needed to be repeated to reach the same result. The effectiveness of the combined entomopathogen application and of the application of S. carpocapsae alone was not influenced by the age of the larvae. However, the single applications of B. bassiana were only effective on young larvae. In both trials, web production of surviving larvae was very significantly reduced by the application of S. carpocapsae in single and combined treatments. CONCLUSIONS: As the results show, only one application is necessary in the combined application of the two entomopathogens to control I. dea, which implies that such treatment is probably the economically most relevant solution.

[Influences of Solenopsis invicta buren invasion on the native ant communities in different habitats in Guangdong].

By using pitfall and bait traps, an investigation was made on the diversity and similarity of ant communities in the areas infested and un-infested with Solenopsis invicta Buren in Shenzhen of Guangdong. The results showed that under the invasion of S. invicta, the ant species number in lawn and wasteland reduced obviously, with a decrease of 6 in lawn and 3 in wasteland, and the native dominant ant species in lichee orchard, especially in wasteland and lawn, were replaced by S. invicta. With the infestation of S. invicta, the diversity and evenness of ant communities in wasteland and lawn decreased but the predominance increased obviously, while it was in adverse in lichee orchard. The similarity coefficients of the ant communities between S. invicta infested and un-infested lichee orchard, wasteland and lawn were 0.6316, 0.5882 and 0.2941, respectively.

Hot-water immersion quarantine treatment against Mediterranean fruit fly and Oriental fruit fly (Diptera: Tephritidae) eggs and larvae in litchi and longan fruit exported from Hawaii.

Immersion of litchi fruit in 49 degrees C water for 20 min followed by hydrocooling in ambient (24 +/- 4 degrees C) temperature water for 20 min was tested as a quarantine treatment against potential infestations of Mediterranean fruit fly, Ceratitis capitata (Wiedemann); and oriental fruit fly, Bactrocera dorsalis Hendel, eggs or larvae in Hawaiian litchi, Litchi chinensis Sonnerat. The 49 degrees C hot-water immersion of litchi provided probit 9 (99.9968% mortality with >95% confidence) quarantine security against eggs and first instars. There were no survivors from 15,000 each feeding and nonfeeding Mediterranean fruit fly or oriental fruit fly third instars immersed in a computer-controlled water bath that simulated the litchi seed-surface temperature profile during the 49 degrees C hot-water immersion treatment. Litchi served as the model for longan, Dimocarpus longan Lour., a closely related fruit that is smaller and also has commercial potential for Hawaii. Modified fruit infestation and holding techniques used to obtain adequate estimated treated populations from poor host fruit, such as litchi and longan, are described. Data from these experiments were used to obtain approval of a hot-water immersion quarantine treatment against fruit flies for litchi and longan exported from Hawaii to the U.S. mainland.

[Ecological control effects of Litchi chinensis-Desmodium intortum complex plant ecosystem on litchi pests].

An investigation on the community structure and dynamics of litchi pests and their natural enemies in constructed Litchi chinensis-Desmodium intortum complex plant ecosystem and single L. chinensis ecosystem showed that the total amount of litchi pests in the complex plant ecosystem was 61.27% of that in the single ecosystem in whole year, and only 50.45% in May, the key time for fruit development, which suggested that there was an interaction between D. intortum and L. chinensis. D. intortum and L. chinensis had a few common pests, but many common natural enemies. D. intortum florescence in winter provided shelter and substitutive food for the natural enemies of pests to survive in the extreme environmental conditions in winter. L. chinensis florescence was on the heel of D. intortum florescence, which provided better conditions for the natural enemies to survive and multiply. During florescence and fruit development stages of L. chinensis (from March to June), the predator/prey ratio in complex plant system was 4.22, 2.34, 2.2 and 20.63 times of that in single plant system in March, April, May and June, respectively, indicating the good control effect on pests of L. chinensis.

Cryptococcus bestiolae and Cryptococcus dejecticola, two new yeast species isolated from frass of the litchi fruit borer Conopomorpha sinensis Bradley.

Two new yeast species, Cryptococcus bestiolae and Cryptococcus dejecticola, were discovered in the frass of the litchi fruit borer Conopomorpha sinensis Bradley. The yeasts utilize inositol, hydrolyze urea, produce starch-like substance, and contain CoQ10. Phylogenetic analyses of D1/D2 26S rDNA and internal transcribed spacer (ITS) sequences indicate that the yeasts are closely related to Bullera dendrophila and an undescribed species of Cryptococcus (strain CBS 8507). The two new species differed from each other by 17 nucleotides in the D1/D2 region and by 68 nucleotides in the ITS region. Cryptococcus bestiolae is a sister species to Cryptococcus sp. CBS 8507, from which it differs by eight nucleotides in the D1/D2 region and 59 nucleotides in the ITS region. Cryptococcus dejecticola and B. dendrophila differed by 13 nucleotides in the D1/D2 and 57 nucleotides in the ITS region. Cryptococcus bestiolae and Cr. dejecticola formed with B. dendrophila a well defined clade consisting of insect associated species. The type strain of Cr. bestiolae is TH3.2.59 (=CBS 10118=NRRL Y-27894), and the type strain of Cr. dejecticola is Litch 17 (=CBS 10117=NRRL Y-27898).